Serrahima-Zieger M, Monteil H, Luu B
Biochim Biophys Acta. 1982 Mar 16;679(3):369-75. doi: 10.1016/0005-2728(82)90156-6.
The membrane-bound ATPase activity of Bacillus subtilis was inhibited by dicyclohexylcarbodiimide (DCCD). The DCCD-reactive proteolipid of B. subtilis was extracted, from labelled or untreated membranes containing F1 or depleted of F1, with neutral or acidic chloroform/methanol. Purification of the [14C]DCCD-binding proteolipid was attempted by column chromatography on methylated Sephadex G-50 and on DEAE-cellulose. The maximal amount of DCCD which could be bound to the purified proteolipid was found to exceed the amount bound by the purified proteolipid extracted from membranes labelled with the lowest [14C]DCCD concentration required for maximal inhibition of the membrane-bound ATPase activity. The radioactive protein peaks eluted by gel filtration and ion-exchange chromatography were analysed by urea-SDS polyacrylamide slab gel electrophoresis and autoradiography. Radioactivity was incorporated into two components of Mr 18 000 and 6000 when proteolipid was purified by methylated Sephadex. The 6000 polypeptide was always present, whatever the extraction and purification procedures. However, the 18 000 polypeptide was present in largest quantity only when proteolipid was extracted from membranes containing F1 and purified by methylated Sephadex. When proteolipid was purified on DEAE-cellulose this [14C]DCCD binding component of Mr 18 000 was absent.
枯草芽孢杆菌的膜结合ATP酶活性受到二环己基碳二亚胺（DCCD）的抑制。用中性或酸性氯仿/甲醇从含有F1或不含F1的标记或未处理的膜中提取枯草芽孢杆菌的DCCD反应性蛋白脂质。尝试通过在甲基化葡聚糖凝胶G-50和DEAE-纤维素上进行柱色谱法纯化[14C]DCCD结合蛋白脂质。发现能够与纯化的蛋白脂质结合的DCCD的最大量超过了从用最大抑制膜结合ATP酶活性所需的最低[14C]DCCD浓度标记的膜中提取的纯化蛋白脂质所结合的量。通过尿素-SDS聚丙烯酰胺平板凝胶电泳和放射自显影分析通过凝胶过滤和离子交换色谱洗脱的放射性蛋白峰。当通过甲基化葡聚糖凝胶纯化蛋白脂质时，放射性被掺入Mr 18000和6000的两个组分中。无论提取和纯化程序如何，6000多肽总是存在。然而，只有当从含有F1的膜中提取蛋白脂质并通过甲基化葡聚糖凝胶纯化时，18000多肽才以最大量存在。当在DEAE-纤维素上纯化蛋白脂质时，不存在这种Mr 18000的[14C]DCCD结合组分。
